MICAL is an oxidoreductase that participates in cytoskeleton reorganization via actin disassembly in the presence of NADPH. Although three MICALs (MICAL1, MICAL2 and MICAL3) have been identified in mammals, only the structure of mouse MICAL1 has been reported. Here, the first crystal structure of human MICAL3, which contains the flavin-containing monooxygenase (FMO) and calponin-homology (CH) domains, is reported. MICAL3 has an FAD/NADP-binding Rossmann-fold domain for monooxygenase activity like MICAL1. The FMO and CH domains of both MICAL3 and MICAL1 are highly similar in structure, but superimposition of the two structures shows a different relative position of the CH domain in the asymmetric unit. Based on kinetic analyses, the catalytic efficiency of MICAL3 dramatically increased on adding F-actin only when the CH domain was available. However, this did not occur when two residues, Glu213 and Arg530, were mutated in the FMO and CH domains, respectively. Overall, MICAL3 is structurally highly similar to MICAL1, which suggests that they may adopt the same catalytic mechanism, but the difference in the relative position of the CH domain produces a difference in F-actin substrate specificity.

Aegilops tauschii (Ae. tauschii) is a diploid (2n = 2x = 14) wild grass species, which has been reported as the progenitor of hexaploid wheat (Triticum aestivum) with D-genome. In this study, 68 Ae. tauschii accessions with diverse geographical backgrounds were investigated for their resistance to infection by the leaf rust fungi Puccinia triticina. Two Ae. tauschii accessions that exhibited hyper-resistance to leaf rust at both seedling and adult stages were identified. Utilizing two susceptible Ae. tauschii ecotypes and keumkang, a common Korean wheat cultivar known to be susceptible to leaf rust, as the negative control, further investigations were conducted for understanding the mechanism underlying immunity to leaf rust disease of these two resistant accessions. Resistant accessions displayed the increased β-1,3-glucanase activity to prevent fungal penetration and the better peroxidase activity to cope with leaf rustinduced oxidative stress. Moreover, transcriptional analyses reveal the important role of the LRR receptor-like serine/ threonine-protein kinase FLS2 (lrr) to the disease resistance of the two ecotypes. Ae. tauschii is a remarkable genetic source, especially for abiotic and biotic stress resistance genes, as the plant is known for its wide-ranging geographical habitat and adaptability to different environments. This, combined with the fact that Ae. tauschii and wheat share a close evolutionary relationship, is indicative of the immense benefit of using Ae. tauschii as a material for improving the quality of synthetic wheat. Our aim was to identify and evaluate the strongest Ae. tauschii contenders for breeding leaf rust-resistant synthetic wheat.

Accumulation of oxidized amino acids, including methionine, has been implicated in aging. The ability to reduce one of the products of methionine oxidation, free methionine-R-sulfoxide (Met-R-SO), is widespread in microorganisms, but during evolution this function, conferred by the enzyme fRMsr, was lost in metazoa. We examined whether restoration of the fRMsr function in an animal can alleviate the consequences of methionine oxidation. Ectopic expression of yeast fRMsr supported the ability of Drosophila to catalyze free Met-R-SO reduction without affecting fecundity, food consumption, and response to starvation. fRMsr expression also increased resistance to oxidative stress. Moreover, it extended lifespan of flies in a methionine-dependent manner. Thus, expression of an oxidoreductase lost during evolution can enhance metabolic and redox functions and lead to an increase in lifespan in an animal model. More broadly, our study exposes the potential of a combination of genetic and nutritional strategies in lifespan control.

Wheat leaf rust caused by Puccinia triticina is a destructive fungal disease causing considerable grain yield loss. In this study, we developed a novel assay to test the rust resistance of detached wheat leaves on defined media with retarded senescence. We observed that salicylic and jasmonic acid confer leaf rust resistance to a susceptible Keumkang wheat (Triticum aestivium L.). Transcription analysis revealed that atchi8 was highly expressed with an increased chitinase activity in the salicylic acid-treated leaves, while expression of PR-9, atpodL, and PR-5 increased in the jasmonic acid-treated leaves. Additionally, the metabolic profile suggested that the phenylalanine pathway might link flavonoid production to leaf rust resistance in the salicylic acid-treated leaves, while the alanine, aspartate, and glutamate metabolism might control the production of other amino acids to enhance pathogen stress response in the jasmonic acid-treated leaves. Finally, all identified genes and metabolites could be potential targets for screening chemical compounds for leaf rust resistance. Future studies on the underlying mechanisms of leaf rust resistance obtained by exogenous treatment of salicylic and jasmonic acids remain necessary.

Methionine oxidation is involved in regulating the protein activity and often leads to protein malfunction. However, tools for quantitative analyses of protein-specific methionine oxidation are currently unavailable. In this work, we developed a biological sensor that quantifies oxidized methionine in the form of methionine-R-sulfoxide in target proteins. The biosensor "tpMe-tROG" consists of methionine sulfoxide reductase B (MsrB), circularly permuted yellow fluorescent protein (cpYFP), thioredoxin, and protein G. Protein G binds to the constant region of antibodies against target proteins, specifically capturing them. Then, MsrB reduces the oxidized methionine in these proteins, leading to cpYFP fluorescence changes. We assessed this biosensor for quantitative analysis of methionine-R-sulfoxide in various proteins, such as calmodulin, IDLO, LegP, Sacde, and actin. We further developed an immunosorbent assay using the biosensor to quantify methionine oxidation in specific proteins such as calmodulin in animal tissues. The biosensor-linked immunosorbent assay proves to be an indispensable tool for detecting methionine oxidation in a protein-specific manner. This is a versatile tool for studying the redox biology of methionine oxidation in proteins.

Leaf rust is the most widespread and destructive fungal disease, and outbreaks have always caused considerable losses in wheat yields. Thus, worldwide increases in wheat production depend on the development of leaf rust-resistant wheat varieties. In this study, we evaluated the resistance of forty Korean wheat cultivars to leaf rust at the seedling stage. Only two Korean wheats, Ol and Jonong, were resistant to leaf rust, whereas the remaining thirty-eight Korean wheats were susceptible to leaf rust. The Ol and Jonong varieties presented larger dry seed weights and higher antioxidant activity in response to leaf rust than the susceptible wheat varieties. No differences in β-1,3-glucanase activity or chlorophyll content between resistant and susceptible wheat varieties were observed. Overall, these results are important for the development of wheat varieties that are highly resistant to leaf rust and to understand the underlying mechanisms that confer leaf rust resistance.